Comparison of internal process control viruses for detection of food and waterborne viruses
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Enteric viruses are pathogens associated with food- and waterborne outbreaks. The recovery of viruses from food or water samples is affected by the procedures applied to detect and concentrate them. The incorporation of an internal process control virus to the analyses allows monitoring the performance of the methodology. The aim of this study was to produce a recombinant adenovirus (rAdV) and apply it together with bacteriophage PP7 as process controls. The rAdV carries a DNA construction in its genome to differentiate it from wild-type adenovirus by qPCR. The stability of both control viruses was evaluated at different pH conditions. The rAdV was stable at pH 3, 7, and 10 for 18 h. PP7 infectious particles were stable at pH 7 and showed a 2.14 log reduction at pH 10 and total decay at pH 3 after 18 h. Three virus concentration methods were evaluated: hollow-fiber tap water ultrafiltration, wastewater ultracentrifugation, and elution-PEG precipitation from lettuce. Total and infectious viruses were quantified and their recoveries were calculated. Virus recovery for rAdV and PP7 by ultrafiltration showed a wide range (2.10–84.42 and 13.54–84.62%, respectively), whereas that by ultracentrifugation was 5.05–13.71 and 6.98–13.27%, respectively. The performance of ultracentrifugation to concentrate norovirus and enteroviruses present in sewage was not significantly different to the recovery of control viruses. For detection of viruses from lettuce, genomic copies of PP7 were significantly more highly recovered than adenovirus (14.74–18.82 and 0.00–3.44%, respectively). The recovery of infectious virus particles was significantly affected during sewage ultracentrifugation and concentration from lettuce. The simultaneous use of virus controls with dissimilar characteristics and behaviors might resemble different enteric viruses.
KeywordsInternal process control virus Hollow fiber ultrafiltration Ultracentrifugation Lettuce Sewage Virus recovery
This work was supported by grants from Universidad de Buenos Aires (SECyT-UBA 20020100100405) and Agencia Nacional de Promoción Científica y Tecnológica (ANPCyT; PICT 2012-2679), Argentina. Funding sources had no involvement in the study design, collection, analysis and interpretation of data, writing of the paper, or the decision to submit the article for publication. We thank Drs. Juan Stupka, Karina Gomes, Daniel Cisterna, and Leila Martínez from the Virology Department of the Instituto Nacional de Enfermedades Infecciosas - ANLIS Dr. Carlos G. Malbrán for their help with norovirus and enterovirus quantitation. We thank María Victoria Gonzalez Eusevi for correction of the English language.
Compliance with ethical standards
This article does not contain any studies with human participants or animals performed by any of the authors.
Conflict of interest
The authors declare that they have no conflict of interest.
- Beer KD, Gargano JW, Roberts VA, Hill VR, Garrison LE, Kutty PK, Hilborn ED, Wade TJ, Fullerton KE, Yoder JS (2015) Outbreaks associated with environmental and undetermined water exposures—United States, 2011–2012. Morb Mortal Wkly Rep 64:842–848. doi: 10.1136/injuryprev-2011-040066 CrossRefGoogle Scholar
- Brandão MLL, Almeida DO, Bispo FCP, Bricio SML, Marin VA, Miagostovich MP (2014) Assessment of microbiological contamination of fresh, minimally processed, and ready-to-eat lettuces (Lactuca sativa), Rio de Janeiro State, Brazil. J Food Sci 79:M961–M966. doi: 10.1111/1750-3841.12459 CrossRefPubMedGoogle Scholar
- Clokie M, Kropinski AM (2001) Bacteriophages: methods and protocols.Google Scholar
- Costafreda MI, Bosch A, Pintó RM (2006) Development, evaluation, and standardization of a real-time TaqMan reverse transcription-PCR assay for quantification of hepatitis A virus in clinical and shellfish samples. Appl Environ Microbiol 72:3846–3855. doi: 10.1128/AEM.02660-05 CrossRefPubMedPubMedCentralGoogle Scholar
- Eurosurveillance Editorial Team (2014) Hepatitis A outbreak: report on results from food trace-back investigation. Euro Surveill 36:20896Google Scholar
- FAO/WHO (2008) Microbiological hazards in fresh fruits and vegetables: meeting report.Google Scholar
- Hata A, Katayama H, Kojima K, Sano S, Kasuga I, Kitajima M, Furumai H (2014) Effects of rainfall events on the occurrence and detection efficiency of viruses in river water impacted by combined sewer overflows. Sci Total Environ 468–469:757–763. doi: 10.1016/j.scitotenv.2013.08.093 CrossRefPubMedGoogle Scholar
- Hill VR, Kahler AM, Jothikumar N, Johnson TB, Hahn D, Cromeans TL (2007) Multistate evaluation of an ultrafiltration-based procedure for simultaneous recovery of enteric microbes in 100-liter tap water samples. Appl Environ Microbiol 73:4218–4225. doi: 10.1128/AEM.02713-06 CrossRefPubMedPubMedCentralGoogle Scholar
- Jothikumar N, Lowther JA, Henshilwood K, Lees DN, Hill VR, Vinje J (2005c) Rapid and sensitive detection of noroviruses by using TaqMan-based one-step reverse transcription-PCR assays and application to naturally contaminated shellfish samples. Appl Environ Microbiol 71:1870–1875. doi: 10.1128/AEM.71.4.1870 CrossRefPubMedPubMedCentralGoogle Scholar
- Kilpatrick DR, Yang CF, Ching K, Vincent A, Iber J, Campagnoli R, Mandelbaum M, De L, Yang SJ, Nix A, Kew OM (2009) Rapid group-, serotype-, and vaccine strain-specific identification of poliovirus isolates by real-time reverse transcription-PCR using degenerate primers and probes containing deoxyinosine residues. J Clin Microbiol 47:1939–1941. doi: 10.1128/JCM.00702-09 CrossRefPubMedPubMedCentralGoogle Scholar
- Matsushita T, Shirasaki N, Tatsuki Y, Matsui Y (2013) Investigating norovirus removal by microfiltration, ultrafiltration, and precoagulation-microfiltration processes using recombinant norovirus virus-like particles and real-time immuno-PCR. Water Res 47:5819–5827. doi: 10.1016/j.watres.2013.07.004 CrossRefPubMedGoogle Scholar
- Monpoeho S, Maul A, Schwartzbrod L, Billaudel S, Virologie LD, Dieu CHUH, Processing D (2001) Best viral elution method available for quantification of enteroviruses in sludge by both cell culture and reverse transcription-PCR. Appl Environ Microbiol 67:2484–2488. doi: 10.1128/AEM.67.6.2484 CrossRefPubMedPubMedCentralGoogle Scholar
- Poma HR, Rajal VB, Blanco Fernández MD, Barril PA, Giordano MO, Masachessi G, Martínez LC, Isa MB, Freire MC, López Riviello G, Cisterna D, Nates SV, Mbayed VA (2013) Evaluation of concentration efficiency of the Pseudomonas aeruginosa phage PP7 in various water matrixes by different methods. Environ Monit Assess 185:2565–2576. doi: 10.1007/s10661-012-2731-9 CrossRefPubMedGoogle Scholar
- Rajal VB, McSwain BS, Thompson DE, Leutenegger CM, Kildare BJ, Wuertz S (2007) Validation of hollow fiber ultrafiltration and real-time PCR using bacteriophage PP7 as surrogate for the quantification of viruses from water samples. Water Res 41:1411–1422. doi: 10.1016/j.watres.2006.12.034 CrossRefPubMedGoogle Scholar